HK1160129A1 - P38 map kinase inhibitors - Google Patents

Abstract

There are provided inter alia compounds of formula (I) wherein R1, Ar, L, X, R3 and Q are as defined in the specification for use in therapy, especially in the treatment of inflammatory diseases.

Description

Field of the invention

The invention relates to compounds which are inhibitors of p38 mitogen-activated protein kinase enzymes (referred to herein as p38 MAP kinase inhibitors), particularly the alpha and gamma kinase sub-types thereof, and their use in therapy, including in pharmaceutical combinations, especially in the treatment of inflammatory diseases, including inflammatory diseases of the lung, such as COPD.

Background of the invention

Four p38 MAPK isoforms (alpha, beta, gamma and delta respectively) have been identified, each displaying a tissue-specific expression pattern. The p38 MAPK alpha and beta isoforms are ubiquitously expressed throughout the body and are found in many different cell types. The p38 MAPK alpha and beta isoforms are inhibited by certain known small molecule p38 MAPK inhibitors. Earlier generations of compounds were highly toxic due to the ubiquitous expression pattern of these isoforms and off-target effects of the compounds. More recent inhibitors are improved to be highly selective for p38 MAPK alpha and beta isoforms and have a wider safety margin.

Less is known about the p38 MAPK gamma and delta isoforms. These isoforms are expressed in specific tissues/cells (unlike the p38 alpha and p38 beta isoforms). The p38 MAPK-delta isoform is expressed more in the pancreas, testes, lung, small intestine and kidney. It is also abundant in macrophages (Smith, S.J. (2006) Br. J. Pharmacol. 149:393-404) and detectable in neutrophils, CD4+ T cells and endothelial cells (www.genecard.org, Hale, K. K. (1999) J. Immunol. 62(7):4246-4252). Very little is known about the expression of p38 MAPK gamma but it is expressed more in brain and heart, as well as in lymphocytes and macrophages (www.genecard.org).

Selective small molecule inhibitors of p38 MAPK-gamma and -delta are not currently available, but one existing compound, BIRB 796, is known to have pan-isoform inhibitory activity. The p38 gamma and p38 delta inhibition is observed at higher concentrations of the compound than those required to inhibit p38 alpha and p38 beta (Kuma, Y. (2005) J. Biol. Chem. 280:19472-19479). BIRB 796 also impaired the phosphorylation of p38 MAPKs or JNKs by the upstream kinase MKK6 or MKK4. Kuma discussed the possibility that the conformational change caused by the binding of the inhibitor to the MAPK may affect the structure of both its phosphorylation site and the docking site for the upstream activator, therefore impairing the phosphorylation of p38 MAPKs or JNKs.

p38 MAP kinase is believed to play a pivotal role in many of the signalling pathways that are involved in initiating and maintaining chronic, persistent inflammation in human disease, for example, severe asthma and COPD. There is now an abundant literature which demonstrates that p38 MAP kinase is activated by a range of pro-inflammatory cytokines and that its activation results in the recruitment and release of further pro-inflammatory cytokines. Indeed, data from some clinical studies demonstrate beneficial changes in disease activity in patients during treatment with p38 MAP kinase inhibitors. For instance Smith, S. J. (2006) Br. J. Pharmacol. 149:393-404 describes the inhibitory effect of p38 MAP kinase inhibitors on cytokine release from human macrophages. Use of inhibitors of p38 MAP kinase in the treatment of chronic obstructive pulmonary disease (COPD) is proposed. Small molecule inhibitors targeted to p38 MAPKα/β have proved to be effective in reducing various parameters of inflammation in cells and tissues obtained from patients with COPD, who are generally corticosteroid insensitive, (Smith, S. J. (2006) Br. J. Pharmacol. 149:393-404) and in vivo animal models (Underwood, D. C. et al. (2000) 279:895-902; Nath, P. et al. (2006) Eur. J. Pharmacol. 544:160-167). Irusen and colleagues also suggested the possibility of involvement of p38 MAPKα/β on corticosteroid insensitivity via reduction of binding affinity of glucocorticoid receptor (GR) in nuclei (Irusen, E. et al. (2002) J. Allergy Clin. Immunol., 109:649-657). Clinical experience with a range of p38 MAP kinase inhibitors, including AMG548, BIRB 796, VX702, SCIO469 and SCIO323 is described in Lee et al. (2005) Current Med. Chem. 12:2979-2994.

COPD is a condition in which the underlying inflammation has been reported to be substantially resistant to the anti-inflammatory effects of inhaled corticosteroids. Consequently, an effective strategy for treating COPD may well be to develop an intervention which both has inherent anti-inflammatory effects and is able to increase the sensitivity of lung tissues from COPD patients to inhaled corticosteroids. The recent publication of Mercado et al. (2007; American Thoracic Society Abstract A56 ) demonstrates that silencing p38 gamma has the potential to restore sensitivity to corticosteroids. Thus there may be a "two pronged" benefit to the use of a p38 MAP kinase inhibitor for the treatment of COPD.

However, the major obstacle hindering the utility of p38 MAP kinase inhibitors in the treatment of human chronic inflammatory diseases has been the toxicity observed in patients. This has been sufficiently severe to result in the withdrawal from clinical development of many of the compounds progressed, including all those specially mentioned above.

International Patent Application No. WO03/072569 discloses 1-phenyl-1H-pyrazol-5-yl ureido derivatives and their use for the treatment of cytokine mediated diseases.

There remains a need to identify and develop new compounds therapeutically useful as p38 MAP kinase inhibitors which have improved therapeutic potential, in particular which are more efficacious, longer acting and/or less toxic at the relevant therapeutic dose. An objective of the present invention is to provide compounds which inhibit p38 MAP kinase with certain sub-type specificity, which show good anti-inflammatory potential.

Summary of the invention

According to the invention, there is provided a compound of formula (I)

• wherein R1 is C1-6 alkyl optionally substituted by a hydroxyl group;
• R2 is H or C1-6 alkyl optionally substituted by a hydroxyl group;
• R3 is H;
• Ar is a naphthyl or a phenyl ring either of which may be optionally substituted by one or more groups independently selected from C1-6 alkyl, C1-6 alkoxy, amino, and C1-4 mono or di-alkyl amino;
• L represents -O(CH2)m- in which m, is 0, 1 or 2;
• Q is selected from: a) a saturated or unsaturated, branched or unbranched C1-10 alkyl chain, wherein at least one carbon (for example 1, 2 or 3 carbons, suitably 1 or 2, in particular 1) is replaced by a heteroatom selected from O, N, and S(O)p, wherein said chain is optionally, substituted by one or more groups independently selected from oxo, halogen, an aryl group, a heteroaryl group and a heterocyclyl group, each aryl, heteroaryl or heterocyclyl group bearing 0 to 3 substituents selected from halogen, C1-6 alkyl, C1-6 alkoxy, C1-6 haloalkyl, amino, and C1-4 mono or dialkyl amino, with the proviso that the atom linked directly to the carbonyl in -NR3C(O)- is not an oxygen or a sulfur atom; andb) a C0-8 alkyl C5-6 heterocycle said heterocyclyl group comprising at least one heteroatom (for example 1, 2 or 3, suitably 1 or 2, in particular 1 heteroatom) selected from O, N and S, and is optionally substituted by one or two or three groups independently selected from halogen, C1-6 alkyl, C1-6 alkoxy, C1-6 haloalkyl, amino, and C1-4 mono or di-alkyl amino; and p is 0, 1 or 2;

or a pharmaceutically acceptable salt or solvate thereof, including all stereoisomers and tautomers thereof. Brief description of the figures

• Figure 1 shows pre-dose time against neutrophil number in BALF for the compound of Example 1 in the LPS-induced neutrophil accumulation test.
• Figure 2 shows pre-dose time against % inhibition of neutrophilia for the compound of Example 1 in the LPS-induced neutrophil accumulation test.

Detailed description of the invention

Alkyl as used herein refers to straight chain or branched chain alkyl, such as, without limitation, methyl, ethyl, propyl, iso-propyl, butyl, and tert-butyl. In one embodiment alkyl refers to straight chain alkyl.

Alkoxy as used herein refers to straight or branched chain alkoxy, for example methoxy, ethoxy, propoxy, butoxy. Alkoxy as employed herein also extends to embodiments in which the oxygen atom is located within the alkyl chain, for example -CH₂CH₂OCH₃ or -CH₂OCH₃. In one embodiment the alkoxy is linked through oxygen to the remainder of the molecule. In one embodiment the disclosure relates to straight chain alkoxy.

In one embodiment of the disclosure there is provided compounds of formula (I), wherein R¹ is methyl, ethyl, propyl, iso-propyl, butyl or tert-butyl, in particular tert-butyl.

In one embodiment R¹ is -C(CH₃)₂CH₂OH.

In one embodiment R² is methyl, ethyl, propyl, iso-propyl, butyl, or tert-butyl, in particular methyl.

In one embodiment R² is -CH₂OH.

In one embodiment R² is in the 2, 3, or 4 position (i.e. ortho, meta or para position), in particular the para (4) position.

In one embodiment Ar is naphthyl.

In one embodiment Ar is not substituted with optional substituents.

In one embodiment Ar is substituted with 1 or 2 groups.

In one embodiment L is -O(CH₂)_m wherein m is 1 or 2.

Halogen includes fluoro, chloro, bromo or iodo, in particular fluoro, chloro or bromo, especially fluoro or chloro.

In one embodiment of the fragment Q it is a C_1-10 alkyl chain wherein at least one carbon (for example 1, 2 or 3 carbons, in particular 1 or 2 carbons) is replaced by a heteroatom selected from O, N, and S(O)p, wherein said chain is optionally, substituted by one or more groups selected from oxo, halogen, an aryl group, a heteroaryl group and a heterocyclyl group, each aryl, heteroaryl or heterocyclyl group bearing 0 to 3 substituents selected from halogen, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, amino, and C_1-4 mono or di-alkyl amino.

In one embodiment Q is a saturated or unsaturated, branched or unbranched C_1-8 alkyl chain or C_1-6 alkyl chain, wherein at least one carbon is replaced by a heteroatom selected from O, N, and S(O)_p.

In one embodiment the heteroatom(s) replacing carbon(s) in the alkyl chain fragment of Q is/are selected from N and O.

In one embodiment the alkyl chain fragment of Q does not bear any optional substituents.

In one embodiment the alkyl chain fragment of Q bears 1, 2, or 3, for example 1 or 2, in particular 1 optional substituent.

It will be clear to persons skilled in the art that the heteroatom may replace a primary, secondary or tertiary carbon, that is a CH₃, -CH₂- or a -CH-, group, as technically appropriate.

In one embodiment p is 0 or 2.

In one embodiment compounds of the disclosure include those in which the fragment -NR³C(O)Q in formula (I) is represented by:         -NR³C(O)CH₂OC_1-6 alkyl, in particular -NHC(O)CH₂OCH₃;         -NHC(O)CH₂O(CH₂)₂OCH₃;         -NHC(O)CH(CH₃)OCH₃;         -NHC(O)CH₂NHCH₃;         -NHC(O)CH₂NH(CH₂)₂OCH₃;         -NHC(O)CH₂SCH₃;         -NHC(O)CH₂S(O)₂CH₃;         -NHC(O)NH₂;         -NR³C(O)NHC_1-7 alkyl, in particular -NHC(O)NHCH₃;         -NHC(O)N(CH₃)₂; or         -NHC(O)CHN[(CH₂)₂OCH₃]₂.

Thus in one embodiment a nitrogen atom in the alkyl chain is directly bonded to the carbonyl of the fragment -NR³C(O) and additionally may, for example, be a terminal amino group, suitably -NR³C(O)N(CH₃)₂ or-NR³C(O)NHCH₃.

In one embodiment Q is a saturated or unsaturated, branched or unbranched C_1-10 alkyl chain, wherein at least one carbon is replaced by a heteroatom selected from O, N, and S(O)p, wherein said chain is substituted by an aryl group bearing 0 to 3 substituents, for example 1, 2 or 3, such as 1 or 2 substituents independently selected from halogen, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, amino and C_1-4 mono or di-alkyl amino. In one embodiment the said aryl group is phenyl, for example substituted phenyl or unsubstituted phenyl.

Examples of the fragment -NR³C(O)Q wherein Q comprises substituted phenyl include: -NR³C(O)CH₂NHCH₂C₆H₅(OCH₃) and -NR³C(O)CH₂N(CH₃)CH₂C₆H₅(OCH₃).

In one embodiment Q is a saturated or unsaturated, branched or unbranched C_1-10 alkyl chain, wherein at least one carbon is replaced by a heteroatom selected from O, N, and S(O)p, wherein said chain is substituted by a heteroaryl group bearing 0 to 3 substituents for example 1, 2 or 3, such as 1 or 2 substituents selected from halogen, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 alkyl amino, and C_1-4 mono or di-alkyl amino. In one embodiment the said heteroaryl group is selected from, thiophene, oxazole, thiazole, isothiazole, imidazole, pyrazole, isoxazole, isothiazole, oxadiazole, 1,2,3 or 1,2,4 triazole, pyridine, pyridazine, pyrimidine, and pyrazine and, in particular pyridine and pyrimidine, especially pyridine.

In one embodiment Q is a saturated or unsaturated, branched or unbranched C_1-10 alkyl chain, wherein at least one carbon is replaced by a heteroatom selected from O, N and S(O)p wherein said chain is substituted by a heterocyclyl group bearing 0 to 3 substituents, for example 1, 2 or 3, such as 1 or 2 substituents selected from halogen, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, amino, and C_1-4 mono or di-alkyl amino.

In one embodiment said heterocyclyl is selected, from a 5 or 6 membered saturated or partially unsaturated ring system comprising one or more (for example 1, 2 or 3 suitably 1 or 2, in particular 1) heteroatom(s) independently selected from O, N and S, for example pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, morpholine, and 1,4-dioxane.

In one embodiment compounds of the disclosure include compounds of formula (I) in which the fragment -NR³C(O)C_0-8 alkylheterocyclyl is represented by:         -NHC(O)-(tetrahydropyranyl), such as -NHC(O)-(tetrahydro-2H-pyran-4-yl),         -NHC(O)-(morpholinyl), such as -NHC(O)-(4-morpholinyl),         -NHC(O)-(pyrrolidinyl), such as -NHC(O)-(pyrrolidin-1-yl),         -NHC(O)-(piperazinyl), such as -NHC(O)-(piperazin-1-yl),         -NHC(O)-(methylpiperazinyl), such as -NHC(O)-(4-methylpiperazin-1-yl),         -NHC(O)-[(methoxyethyl)piperazinyl], such as -NHC(O)-[4-(2-methoxyethyl)piperazin-1-yl],         -NHC(O)CH₂-(tetrahydropyranyl), such as -NHC(O)CH₂-(tetrahydro-2H-pyran-4-yl),         -NHC(O)CH₂-(morpholinyl), such as -NHC(O)CH₂-(4-morpholinyl),         -NHC(O)CH₂-(pyrrolidinyl), such as -NHC(O)CH₂-(pyrrolidin-1-yl),         -NHC(O)CH₂-(piperazinyl), such as -NHC(O)CH₂-(piperazin-1-yl),         -NHC(O)CH₂-(methylpiperazinyl), such as -NHC(O)CH₂-(4-methylpiperazin-1-yl), or         -NHC(O)CH₂-[(methoxyethyl)piperazinyl], such as -NHC(O)CH₂-[4-(2-methoxyethyl)piperazin-1-yl].

In one embodiment of the fragment Q, the saturated or unsaturated, branched or unbranched C_1-10 alkyl chain, wherein at least one carbon is replaced by a heteroatom selected from O, N, and S(O)_p is selected from: -CH₂OCH₂-, -CH₂NHCH₂-, -CH₂NH- and -CH₂OCH₂CH₂-. These fragments may optionally terminate in an aryl group, a heteroaryl group or a heterocyclyl group as defined for fragment Q above.

In one embodiment the disclosure relates to compounds of formula (IA) wherein R¹, R², Ar, L and R³ are as defined above and p is 0, 1 or 2, in particular 0 or 2, especially 0, and x is an integer from 1 to 6 (including 2, 3, 4 and 5) and y is zero or an integer from 1 to 5 (including 2, 3 and 4) with the proviso that x + y is an integer from 1 to 6, for example x is 1 and y is 1.

In another embodiment the disclosure relates to compounds of formula (IB)

• wherein R1, R2, Ar, L and R3 are as defined above,
• x is an integer from 1 to 6 (including 2, 3, 4 and 5) and y is zero or an integer from 1 to 5 (including 2, 3 and 4),

with the proviso that x + y is an integer from 1 to 6, for example x is 1 and y is 0.

In one embodiment of the compounds of formula (IB) the fragment -NR³C(O)(CH₂)_xO(CH₂)_yCH₃ is: -NHC(O)CH₂OCH₃.

In one embodiment the compound is not: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxyacetamide.

In one embodiment the compound is:

• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxyacetamide;
• 1-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(3-methylureido)pyridine-4-yl)methoxy)naphthalene-1-yl)urea;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)tetrahydro-2H-pyran-4-carboxamide;
• (S)-N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxypropanamide;
• (R)-N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxypropanamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(methylthio)acetamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-morpholinoacetamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)u reido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(pyrrolidin-1-yl)acetamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(4-methylpiperazin-1-yl)acetamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(4-(2-methoxyethyl)piperazin-1-y)acetamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(2-methoxyethylamino)acetamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(dimethylamino)acetamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(methylamino)acetamide;
• N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-((4-methoxybenzyl)(methyl)amino)acetamide;
• N-(4-(2-(4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)ethyl)pyridin-2-yl)-2-methoxyacetamide;
• N-(4-(2-(4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)ethyl)pyridin-2-yl)-2-(2-methoxyethoxy)acetamide;
• 1-(3-(tert-Butyl)-1-(p-tolyl)-1 H-pyrazol-5-yl)-3-(4-(2-(2-(3-methylureido)pyridine-4-yl)ethoxy)naphthalen-1-ylurea; or
• 1-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-(2-(2-ureidopyridine-4-yl)ethoxy)naphthalen-1-ylurea;

or a pharmaceutically acceptable salt or solvate thereof, including all stereoisomers and tautomers thereof.

Examples of salts of compounds of formula (I) include all pharmaceutically acceptable salts, such as, without limitation, acid addition salts of strong mineral acids such as HCl and HBr salts and addition salts of strong organic acids such as a methansulfonic acid salt.

Examples of solvates include hydrates.

The compounds described herein may include one or more chiral centres, and the disclosure extends to include racemates, enantiomers and stereoisomers resulting therefrom. In one embodiment one enantiomeric form is present in a substantially purified form that is substantially free of the corresponding entaniomeric form.

The disclosure also extends to all polymorphic forms of the compounds herein defined.

Compounds of formula (I) can be prepared by a process comprising reacting compounds of formula (II): where Ar, L, R¹, R² and R³ are as defined above for compounds of formula (I) with a compound of formula (III): where Q is as defined above for compounds of formula (I), and LG₁ is a leaving group for example halogen, such as chloro.

The reaction is suitably carried out in the presence of a base (e.g. diisopropylethylamine). The reaction is suitably carried out in an aprotic solvent or solvent mixture, e.g. DCM and DMF.

Compounds of formula (II) can be prepared by reacting a compound of formula (IV) where R¹ and R² are as defined above for compounds of formula (I), with a compound of formula (V):

• wherein Ar, L and R3 are defined above for compounds of formula (I)

and a compound of formula (VI): wherein LG₂ and LG₃ each independently represent leaving groups (e.g. LG₂ and LG₃ both represent imidazolyl or halogen such as chloro).

The reaction is suitably carried out in an aprotic solvent (e.g. dichloromethane), using appropriate protecting groups for chemically sensitive groups.

A compound of formula (V) may be prepared by reduction of a compound of formula (VII) wherein Ar, L and R³ are as defined above for compounds of formula (I), for example by hydrogenation in the presence of a catalyst such as platinum supported on carbon.

The reaction is suitably carried out in polar protic solvent or mixture of solvents (e.g. methanol and acetic acid).

A compound of formula (VII) may be prepared by reaction of a compound of formula (VIII) wherein R³ and m are as defined above for formula (I) with a compound of formula (IX) or (X) wherein compounds (IX) and (X) may bear optional substitutents as defined above for compounds of formula (I).

The reaction may be performed under Mitsunobu conditions, such as in the presence of triphenylphosphine and diisopropylazodicarboxylate. The reaction is suitably carried out in a polar aprotic solvent (e.g. tetrahydrofuran, in particular anhydrous tetrahydrofuran).

Compounds of formulae (III), (IV), (VI), (VIII), (IX) and (X) are either commercially available, or are known, or are novel and can be readily prepared by conventional methods. See for example Regan, J. et al.; J. Med. Chem., 2003, 46, 4676-4686, WO00/043384 , WO2007/087448 and WO2007/089512 .

Protecting groups may be required to protect chemically sensitive groups during one or more of the reactions described above, to ensure that the process is efficient. Thus if desired or necessary, intermediate compounds may be protected by the use of conventional protecting groups. Protecting groups and means for their removal are described in "Protective Groups in Organic Synthesis", by Theodora W. Greene and Peter G.M. Wuts, published by John Wiley & Sons Inc; 4th Rev Ed., 2006, ISBN-10: 0471697540.

Novel intermediates are claimed as an aspect of the invention.

In one aspect the compounds are useful in treatment, for example in treating COPD and/or asthma.

The compounds developed to date have typically been intended for oral administration. This strategy involves optimizing compounds which achieve their duration of action by an appropriate pharmacokinetic profile. This ensures that there is a sufficient drug concentration established and maintained after and between doses to provide clinical benefit. The inevitable consequence of this approach is that all body tissues, especially liver and gut, are likely to be exposed to therapeutically active concentrations of the drug, whether or not they are adversely affected by the disease being treated.

An alternative strategy is to design treatment approaches in which the drug is dosed directly to the inflamed organ (topical therapy). Whilst this approach is not suitable for treating all chronic inflammatory diseases, it has been extensively exploited in lung diseases (asthma, COPD), skin diseases (atopic dermatitis and psoriasis), nasal diseases (allergic rhinitis) and gastrointestinal diseases (ulcerative colitis).

In topical therapy, efficacy can be achieved either by (i) ensuring that the drug has a sustained duration of action and is retained in the relevant organ to minimize the risks of systemic toxicity or (ii) producing a formulation which generates a "reservoir" of the active drug which is available to sustain the drug's desired effects. Approach (i) is exemplified by the anticholinergic drug tiotropium (Spiriva), which is administered topically to the lung as a treatment for COPD, and which has an exceptionally high affinity for its target receptor resulting in a very slow off rate and a consequent sustained duration of action.

In one aspect of the disclosure the compounds herein are particularly suitable for topical delivery, such as topical delivery to the lungs, in particular for the treatment of COPD.

In one aspect the compounds have a longer duration of actions than BIRB 796.

In one embodiment the compounds are suitable for sensitizing patients to treatment with a corticosteroid.

The compounds herein may also be useful for the treatment of rheumatoid arthritis.

Further, the present invention provides a pharmaceutical composition comprising a compound according to the disclosure optionally in combination with one or more pharmaceutically acceptable diluents or carriers.

Diluents and carriers may include those suitable for parenteral, oral, topical, mucosal and rectal administration.

As mentioned above, such compositions may be prepared e.g. for parenteral, subcutaneous, intramuscular, intravenous, intra-articular or peri-articular administration, particularly in the form of liquid solutions or suspensions; for oral administration, particularly in the form of tablets or capsules; for topical e.g. pulmonary or intranasal administration, particularly in the form of powders, nasal drops or aerosols and transdermal administration; for mucosal administration e.g. to buccal, sublingual or vaginal mucosa, and for rectal administration e.g. in the form of a suppository.

The compositions may conveniently be administered in unit dosage form and may be prepared by any of the methods well-known in the pharmaceutical art, for example as described in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, PA., (1985). Formulations for parenteral administration may contain as excipients sterile water or saline, alkylene glycols such as propylene glycol, polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, hydrogenated naphthalenes and the like. Formulations for nasal administration may be solid and may contain excipients, for example, lactose or dextran, or may be aqueous or oily solutions for use in the form of nasal drops or metered sprays. For buccal administration typical excipients include sugars, calcium stearate, magnesium stearate, pregelatinated starch, and the like.

Compositions suitable for oral administration may comprise one or more physiologically compatible carriers and/or excipients and may be in solid or liquid form. Tablets and capsules may be prepared with binding agents, for example, syrup, acacia, gelatin, sorbitol, tragacanth, or poly-vinylpyrollidone; fillers, such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, or glycine; lubricants, such as magnesium stearate, talc, polyethylene glycol, or silica; and surfactants, such as sodium lauryl sulfate. Liquid compositions may contain conventional additives such as suspending agents, for example sorbitol syrup, methyl cellulose, sugar syrup, gelatin, carboxymethyl-cellulose, or edible fats; emulsifying agents such as lecithin, or acacia; vegetable oils such as almond oil, coconut oil, cod liver oil, or peanut oil; preservatives such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). Liquid compositions may be encapsulated in, for example, gelatin to provide a unit dosage form.

Solid oral dosage forms include tablets, two-piece hard shell capsules and soft elastic gelatin (SEG) capsules.

A dry shell formulation typically comprises of about 40% to 60% concentration of gelatin, about a 20% to 30% concentration of plasticizer (such as glycerin, sorbitol or propylene glycol) and about a 30% to 40% concentration of water. Other materials such as preservatives, dyes, opacifiers and flavours also may be present. The liquid fill material comprises a solid drug that has been dissolved, solubilized or dispersed (with suspending agents such as beeswax, hydrogenated castor oil or polyethylene glycol 4000) or a liquid drug in vehicles or combinations of vehicles such as mineral oil, vegetable oils, triglycerides, glycols, polyols and surface-active agents.

Suitably the compound of formula (I) is administered topically to the lung. Hence we provide according to the invention a pharmaceutical composition comprising a compound of the disclosure optionally in combination with one or more topically acceptable diluents or carriers. Topical administration to the lung may be achieved by use of an aerosol formulation. Aerosol formulations typically comprise the active ingredient suspended or dissolved in a suitable aerosol propellant, such as a chlorofluorocarbon (CFC) or a hydrofluorocarbon (HFC). Suitable CFC propellants include trichloromonofluoromethane (propellant 11), dichlorotetrafluoromethane (propellant 114), and dichlorodifluoromethane (propellant 12). Suitable HFC propellants include tetrafluoroethane (HFC-134a) and heptafluoropropane (HFC-227). The propellant typically comprises 40% to 99.5% e.g. 40% to 90% by weight of the total inhalation composition. The formulation may comprise excipients including co-solvents (e.g. ethanol) and surfactants (e.g. lecithin, sorbitan trioleate and the like). Aerosol formulations are packaged in canisters and a suitable dose is delivered by means of a metering valve (e.g. as supplied by Bespak, Valois or 3M).

Topical administration to the lung may also be achieved by use of a non-pressurised formulation such as an aqueous solution or suspension. This may be administered by means of a nebuliser. Topical administration to the lung may also be achieved by use of a dry-powder formulation. A dry powder formulation will contain the compound of the disclosure in finely divided form, typically with a mass mean diameter (MMAD) of 1-10 microns. The formulation will typically contain a topically acceptable diluent such as lactose, usually of large particle size e.g. a mass mean diameter (MMAD) of 100um or more. Example dry powder delivery systems include SPINHALER, DISKHALER, TURBOHALER, DISKUS and CLICKHALER.

Compounds according to the disclosure are intended to have therapeutic activity. In a further aspect, the present invention provides a compound of the disclosure for use as a medicament.

Compounds according to the disclosure may also be useful in the treatment of respiratory disorders including COPD (including chronic bronchitis and emphysema), asthma, paediatric asthma, cystic fibrosis, sarcoidosis, idiopathic pulmonary fibrosis, allergic rhinitis, rhinitis, sinusitis, especially asthma, chronic bronchitis and COPD.

Compounds of the disclosure may also re-sensitise the patient's condition to treatment with a corticosteroid, when the patient's condition has become refractory to the same.

Compounds according to the disclosure are also expected to be useful in the treatment of certain conditions which may be treated by topical or local therapy including allergic conjunctivitis, conjunctivitis, allergic dermatitis, contact dermatitis, psoriasis, ulcerative colitis, inflamed joints secondary to rheumatoid arthritis or osteoarthritis.

Compounds of the disclosure are also expected to be useful in the treatment of certain other conditions including rheumatoid arthritis, pancreatitis, cachexia, inhibition of the growth and metastasis of tumours including non-small cell lung carcinoma, breast carcinoma, gastric carcinoma, colorectal carcinomas and malignant melanoma.

Thus, in a further aspect, the present invention provides a compound as described herein for use in the treatment of the above mentioned conditions.

In a further aspect, the present invention provides use of a compound as described herein for the manufacture of a medicament for the treatment of the above mentioned conditions.

In a further aspect, the present invention provides a method of treatment of the above mentioned conditions which comprises administering to a subject an effective amount of a compound of the disclosure or a pharmaceutical composition thereof.

The word "treatment" is intended to embrace prophylaxis as well as therapeutic treatment.

A compound of the disclosure may also be administered in combination with one or more other active ingredients e.g. active ingredients suitable for treating the above mentioned conditions. For example possible combinations for treatment of respiratory disorders include combinations with steroids (e.g. budesonide, beclomethasone dipropionate, fluticasone propionate, mometasone furoate, fluticasone furoate), beta agonists (e.g. terbutaline, salbutamol, salmeterol, formoterol) and/or xanthines (e.g. theophylline).

Abbreviations

AcOH

glacial acetic acid

aq

aqueous

Ac

acetyl

ATP

adenosine-5'-triphosphate

BALF

bronchoalveolae lavage fluid

br

broad

BSA

bovine serum albumin

CatCart®

catalytic cartridge

CDI

1,1-carbonyl-diimidazole

COPD

chronic obstructive pulmonary disease

d

doublet

DCM

dichloromethane

DIAD

diisopropylazadicarboxylate

DIBAL-H

diisobutylaluminium hydride

DIPEA

N,N-diisopropylethylamine

DMF

N,N-dimethylformamide

DMSO

dimethyl sulfoxide

EtOAc

ethyl acetate

FCS

foetal calf serum

hr

hour(s)

HRP

horseradish peroxidase

JNK

c-Jun N-terminal kinase

MAPK

mitogen protein activated protein kinase

MeOH

methanol

min

minute(s)

MTT

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PBS

phosphate buffered saline

PPh3

triphenylphosphine

RT

room temperature

RP HPLC

reverse phase high performance liquid chromatography

s

singlet

SCX

solid supported cation exchange

SDS

sodium dodecyl sulfate

t

triplet

TFA

trifluoroacetic acid

THF

tetrahydrofuran

TMB

3,3', 5,5'-tetramethylbenzidine

TNFα

tumor necrosis factor alpha

General Procedures

All starting materials and solvents were either obtained from commercial sources or prepared according to the literature citation.

Hydrogenations were preformed on a Thales H-cube flow reactor under the conditions stated.

Organic solutions were routinely dried over magnesium sulfate.

SCX was purchased with Supelco and treated with 1 M aqueous HCl prior to use. The reaction mixture to be purified was first diluted with MeOH and made acidic with a few drops of AcOH. This solution was loaded directly onto the SCX and washed with MeOH. The desired material was then eluted by washing with 1% NH₃ in MeOH.

Column chromatography was performed on Silicycle pre-packed silica (230-400 mesh, 40-63 µm) cartridges using the amount indicated.

Preparative Reverse Phase High Performance Liquid Chromatography:

Agilent Scalar column C18, 5 µm (21.2 x 50 mm), flow rate 28 mL/min eluting with a H₂O-MeCN gradient containing 0.1% v/v formic acid over 10 min using UV detection at 215 and 254 nm. Gradient information: 0.0 - 0.5 min: 95% H₂O-5% MeCN; 0.5 - 7.0 min; Ramped from 95% H₂O-5% MeCN to 5% H₂O-95% MeCN; 7.0 - 7.9 min: Held at 5% H₂O-95% MeCN; 7.9 - 8.0 min: Returned to 95% H₂O-5% MeCN; 8.0 - 10.0 min: Held at 95% H₂O-5% MeCN.

Analytical Methods Reverse Phase High Performance Liquid Chromatography:

Agilent Scalar column C18, 5 µm (4.6 x 50 mm) or Waters XBridge C18, 5 µm (4.6 x 50 mm) flow rate 2.5 mL/min eluting with a H₂O-MeCN gradient containing 0.1% v/v formic acid over 7 min employing UV detection at 215 and 254 nm. Gradient information: 0.0 - 0.1 min: 95% H₂O-5% MeCN; 0.1 -5.0 min; Ramped from 95% H₂O-5% MeCN to 5% H₂O-95% MeCN; 5.0 - 5.5 min: Held at 5% H₂O-95% MeCN; 5.5 - 5.6 min: Held at 5% H₂O-95% MeCN, flow rate increased to 3.5 mL/min; 5.6 - 6.6 min: Held at 5% H₂O-95% MeCN, flow rate 3.5 mL/min; 6.6 - 6.75 min: Returned to 95% H₂O-5% MeCN, flow rate 3.5 mL/min; 6.75 - 6.9 min: Held at 95% H₂O-5% MeCN, flow rate 3.5 mL/min; 6.9 - 7.0 min: Held at 95% H₂O-5% MeCN, flow rate reduced to 2.5 mL/min. ¹H NMR Spectroscopy:

Bruker Avance III 400 MHz using residual undeuterated solvent as reference. Intermediate A: 1-(4-((2-Aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1 H-pyrazol-5-yl)urea

2-Amino-4-((4-nitronaphthalen-1-yloxy)methyl)pyridine (2)

To a solution of 4-nitronaphthol (5.17 g, 27.3 mmol), PPh₃ (10.75 g, 41.0 mmol) and 2-aminopyridine-4-methanol (1) (5.09 g, 41.0 mmol) in THF (50 mL) was added dropwise DIAD (8.07 mL, 41.0 mmol) at -15°C. The mixture was stirred overnight at RT and the volatiles removed in vacuo. The crude product was triturated from EtOAc (150 mL), filtered off and washed with EtOAc (100 mL). A second trituration from MeOH (100 mL) gave 2-amino-4-((4-nitronaphthalen-1-yloxy)methyl)pyridine (2) (4.54 g, 56%) as a yellow solid: m/z 296 (M+H)⁺ (ES⁺).

2-Amino-4-((4-aminonaphthalen-1-yloxy)methyl)pyridine (3)

2-Amino-4-((4-nitronaphthalen-1-yloxy)methyl)pyridine (2) (4.50 g, 15.24 mmol) in MeOH (200 mL) and AcOH (200 mL) was passed through a Thales H-cube (2.0 mL.min^-1, 40°C, 55 mm 10% Pt/C Cat-Cart, full hydrogen mode) and the volatiles were removed in vacuo. The crude product was subjected to SCX capture and release eluting with 1% NH₃ in MeOH solution and the solvent was removed in vacuo to give 2-amino-4-((4-aminonaphthalen-1-yloxy)methyl)pyridine (3) (3.82g, 94%) as a purple solid: m/z 266 (M+H)⁺ (ES⁺).

(Intermediate A): 1-(4-((2-Aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea

To a solution of CDI (4.18 g, 25.8 mmol) in DCM (15 mL) was added dropwise under nitrogen a solution of 3-tert-butyl-1-p-tolyl-1H-pyrazol-5-amine (4) ( WO 2000/043384 ) (5.91g, 25.8 mmol) in DCM (15 mL) over 40 min. The resulting solution was stirred at RT for 1 hr then added dropwise under nitrogen to a solution of 2-amino-4-((4-aminonaphthalen-1-yloxy)methyl)pyridine (3) (3.80 g, 12.9 mmol). The mixture was stirred overnight and the volatiles were removed in vacuo. The crude material was purified by column chromatography (120 g); eluting with 0 to 6% MeOH in DCM to give 1-(4-((2-aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate A) as an off white solid (4.27 g, 63%): m/z 521 (M+H)⁺ (ES⁺).

Example 1: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl) pyridin-2-yl)-2-methoxyacetamide

To a mixture of 1-(4-((2-aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate A) (526 mg, 0.96 mmol) and DIPEA (184 µL, 1.06 mmol) in DCM/DMF (10:1, 11 mL) was added methoxyacetyl chloride (92 µL, 1.01 mmol). After stirring for 1 hr at RT, further DIPEA (184 µL, 1.06 mmol) and methoxyacetyl chloride (92 µL, 1.01 mmol) were added sequentially and stirring was continued for 1 hr. After the addition of a solution of 1% NH₃ in MeOH (40 mL), the mixture was stirred for 15 min and evaporated in vacuo. The crude product was purified by column chromatography (40 g); eluting with 0 to 6% MeOH in DCM to furnish N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxyacetamide (Example 1) as a white solid (286 mg, 49%): m/z 593 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.39 (3H, s), 3.32 (3H, s), 4.08 (2H, s), 5.39 (2H, s), 6.36 (1 H, s), 7.03 (1 H, d), 7.28 (1 H, dd), 7.36 (2H, m), 7.44 (2H, m), 7.56-7.64 (3H, m), 7.93 (1 H, m), 8.30-8.35 (3H, m), 8.58 (1 H, s), 8.79 (1 H, s), 10.02 (1 H, s).

Example 2: 1-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(3-methylureido)pyridine-4-yl)methoxy)naphthalene-1-yl)urea

To a solution of 1-(4-((2-aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate A) (70 mg, 0.13 mmol) in anhydrous pyridine (1.5 mL) was added methyl isocyanate (14 µL, 0.24 mmol) and the mixture allowed to stir at RT for 72 hr. Pyridine was removed under vacuum and the residue triturated with DCM (3.0 mL). Filtration afforded an off-white powder, 1-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(3-methylureido)pyridine-4-yl)methoxy)naphthalene-1-yl)urea (Example 2) (36 mg, 45 %): m/z 578 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.39 (3H, s), 2.74 (3H, d), 5.30 (2H, s), 6.36 (1H, s), 6.99 (1H, d), 7.05 (d, 1 H), 7.35, (2H, d), 7.44 (2H, d), 7.54-7.64 (4H, m), 7.93 (1 H, d), 8.19 (1 H, d), 8.23 (1 H, brs), 8.35 (1 H, d), 8.58 (1 H, s), 8.79 (1 H, s), 9.36 (1 H, s).

Example 3: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl) pyridin-2-yl)tetrahydro-2H-pyran-4-carboxamide

DMF (2 drops) was added to a stirred solution of tetrahydropyran-2H-4-carboxylic acid and oxalyl chloride (21 µL, 0.25 mmol) in DCM (1.0 mL) and the resulting solution was stirred at RT for 1 hr. The solution was evaporated in vacuo to give a colourless oil, which was redissolved in DCM (1.0 mL) and added dropwise to a stirred mixture of 1-(4-((2-aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate A) (50 mg, 0.10 mmol) and DIPEA (84 µL, 0.50 mmol) in DCM (1.0 mL). Stirring was continued for 18 hr. The reaction mixture was stirred in 1% NH₃ in MeOH (20 mL) for 30 min, evaporated in vacuo, pre-adsorbed on silica, and purified by column chromatography (12 g, 0-5% MeOH in DCM, gradient elution) to give N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido) naphthalen-1-yloxy)methyl)pyridin-2-yl)tetrahydro-2H-pyran-4-carboxamide (Example 3) as a light tan solid (18 mg, 28%): m/z 633 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.26 (9H, s), 1.57-1.72 (4H, m), 2.38 (3H, s), 2.75 (1 H, m), 3.28-3.33 (2H, m), 3.88 (2H, m), 5.35 (2H, s), 6.34 (1H, s), 6.99 (1H, d), 7.24 (1H, dd), 7.35 (2H, m), 7.43 (2H, m), 7.55-7.64 (3H, m), 7.92 (1 H, m), 8.27-8.33 (3H, m), 8.58 (1 H, s), 8.78 (1H, s), 10.50 (1 H, s).

Example 4: (S)-N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-methoxypropanamide

1-Chloro-N,N-dimethylethenamine (50 µL, 0.48 mmol) was added to a stirring solution of (S)-2-methoxypropionic acid (50 mg, 0.48 mmol) in DCM (1.0 mL) and the resulting yellow solution was stirred at RT for 1 hr. The solution was added dropwise to a stirring mixture of 1-(4-((2-aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(Intermediate A) (50 mg, 0.10 mmol) and DIPEA (167 µl, 0.96 mmol) in DCM (1.0 mL). Stirring was continued overnight. The reaction mixture was stirred in 1% NH₃ in MeOH (20 mL), evaporated in vacuo, pre-adsorbed on silica and purified by column chromatography (12 g, 10-50% EtOAc in iso-hexane, gradient elution) to give (S)-N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxypropanamide (Example 4) as a colourless solid (18 mg, 30%): m/z 607 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, d), 1.31 (3H, s), 2.38 (3H, s), 3.30 (3H, s), 4.02 (1 H, q), 5.39 (2H, s), 6.37 (1 H, s), 7.00 (1H, d), 7.29 (1 H, dd), 7.35 (2H, m), 7.45 (2H, m), 7.56-7.64 (3H, m), 7.93 (1 H, m), 8.30-8.37 (3H, m), 8.58 (1 H, s), 8.79 (1 H, s), 10.06 (1 H, s).

Example 5: (R)-N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-methoxypropanamide

1-Chloro-N,N-dimethylethenamine (38 µL, 0.36 mmol) was added to a stirred solution of (R)-2-methoxypropionic acid (37 mg, 0.36 mmol) in DCM (1.0 mL) and the resulting solution was stirred at RT for 1 hr. The solution was added dropwise to a stirred mixture of 1-(4-((2-aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate A) (75 mg, 0.14 mmol) and DIPEA (75 µL, 0.43 mmol) in DCM (2.0 mL) at 0°C. Stirring was continued for a further 48 hr. The mixture was poured in to 1% NH₃ in MeOH (20 mL) and stirred for 1 hr, and evaporated in vacuo to give a yellow residue. Column chromatography (12 g, 20-50% EtOAc in iso-hexane) gave (R)-N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxypropanamide (Example 5) as a light pink solid (39 mg, 43%): m/z 607 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, d), 1.30 (3H, s), 2.39 (3H, s), 3.31 (3H, s), 4.02 (1H, q), 5.39 (2H, s), 6.35 (1H, s), 7.02 (1H, d), 7.29 (1H, dd), 7.35 (2H, m), 7.45 (2H, m), 7.56-7.64 (3H, m), 7.93 (1 H, m), 8.30-8.37 (3H, m), 8.58 (1 H, s), 8.79 (1 H, s), 10.09 (1 H, s).

Intermediate B: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-chloroacetamide

To a solution of DIPEA (1.37 mL, 7.68 mmol) and 1-(4-((2-aminopyridin-4-yl)methoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate A) (2.00 g, 3.84 mmol) in DCM (40 mL) and DMF (8.0 mL) was added chloroacetyl chloride (0.61 mL, 7.68 mmol). The reaction mixture was stirred at RT for 1 hr. LC-MS indicated nearly complete consumption of the starting material. A further portion of chloroacetyl chloride (100µl, 1.25mmol) was added. After stirring for 1 hr at RT, the reaction mixture was partitioned between DCM (40 mL) and saturated aq NaHCO₃ solution (40 mL). The organic phase was concentrated in vacuo and purified by column chromatography (80 g, 0-10% MeOH in DCM, gradient elution). Product fractions were concentrated in vacuo and the residue triturated with diethyl ether (20 mL) and iso-hexane (20 mL). The solid was collected by filtration to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) as a light purple solid (1.07 g, 42%): m/z 597, 599 (M+H)⁺ (ES⁺).

Example 6: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl) pyridin-2-yl)-2-(methylthio)acetamide

N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (100 mg, 0.17 mmol) was added portionwise to a stirred mixture of sodium thiomethoxide (35 mg, 0.50 mmol) in MeOH (5.0 mL) and the resulting mixture was stirred for 1 hr at RT. The mixture was evaporated in vacuo and partitioned between brine (20 mL) and DCM (30 mL). The organic layer was concentrated in vacuo, the residue pre-adsorbed on silica and purified by column chromatography (12 g, 10-100% EtOAc in iso-hexane, gradient elution). Product fractions were evaporated in vacuo to give N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(methylthio)acetamide (Example 6) as a light yellow solid (28 mg, 26%): m/z 610 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.16 (3H, s), 2.39 (3H, s), 3.53 (2H, s), 5.37 (2H, s), 6.35 (1 H, s), 7.01 (1 H, d), 7.26 (1 H, dd), 7.35 (2H, m), 7.44 (2H, m), 7.55-7.64 (3H, m), 7.92 (1 H, m), 8.30-8.35 (3H, m), 8.58 (1 H, s), 8.78 (1 H, s), 10.60 (1 H, s).

Example 7: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl) pyridin-2-yl)-2-morpholinoacetamide

To a solution of N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (50 mg, 0.08 mmol) in DCM (1.0 mL), DMF (0.1 mL) and DIPEA (21.9 µL, 0.13 mmol) was added morpholine (11.0 µL, 0.13 mmol). The reaction mixture was stirred at RT for 3 hr. LC-MS indicated 20% conversion to product. The reaction mixture was heated to 40°C and stirred for 12 hr. LC-MS indicated 87% conversion to product. A further portion of morpholine (11.0 µL, 0.13 mmol) was added and the reaction mixture stirred at 40°C for 5 hr. LC-MS indicated 94% conversion to product. The crude reaction mixture was purified by column chromatography (12 g, 0-10% MeOH in DCM, gradient elution). Product fractions were concentrated in vacuo and the residue triturated with MeOH (5.0 mL). The solid was collected by filtration to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-morpholinoacetamide (Example 7) as a light yellow solid (11 mg, 20%): m/z 648 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.39 (3H, s), 2.54 (4H, m), 3.20 (2H, s), 3.63 (4H, m), 5.39 (2H, s), 6.35 (1 H, s), 7.01 (1 H, d), 7.28 (1 H, d), 7.35 (2H, d), 7.43 (2H, d), 7.63 - 7.56 (3H, m), 7.92 (1 H, d), 8.37 - 8.29 (3H, m), 8.58 (1 H, s), 8.79 (1 H, s), 10.01 (1 H, s).

Example 8: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-(pyrrolidin-1-yl)acetamide

To a solution of N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (50 mg, 0.08 mmol) in DCM (1.0 mL), DMF (0.1 mL) and DIPEA (22 µL, 0.13 mmol) was added pyrrolidine (7.0 µL, 0.08 mmol). The reaction mixture was stirred at RT for 3 hr. LC-MS indicated 50% conversion to product. The reaction mixture was heated to 40°C and stirred for 12 hr. LC-MS indicated 95% conversion to product. A further portion of pyrrolidine (7.0 µL, 0.08 mmol) was added and the reaction mixture continued to stir at 40°C for 5 hr. LC-MS indicated complete conversion to product. The crude reaction mixture was purified by column chromatography (12 g, 0-10% MeOH in DCM, gradient elution) to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(pyrrolidin-1-yl)acetamide (Example 8) as a light orange solid (17 mg, 32%): m/z 632 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 1.76 (4H, m), 2.39 (3H, s), 2.62 (4H, m), 5.39 (2H, s), 6.35 (1H, s), 7.01 (1H, d), 7.28 (1H, d), 7.34 (2H, d), 7.44 (2H, d), 7.65 - 7.55 (3H, m), 7.92 (1H, d), 8.36 - 8.29 (3H, m), 8.58 (1H, s), 8.79 (1H, s), 9.93 (1H, s).

Example 9: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl) pyridin-2-yl)-2-(4-methylpiperazin-1-yl)acetamide

To a solution of N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (50 mg, 0.08 mmol) in DCM (1.0 mL), DMF (0.1 mL) and DIPEA (22 µL, 0.13 mmol) was added N-methyl piperazine (9.3 µl, 0.08 mmol). The reaction mixture was stirred at RT for 3 hr. LC-MS indicated 20% conversion to product. The reaction mixture was heated to 40°C and stirred for 12 hr. LC-MS indicated 91% conversion to product. A further portion of N-methyl piperazine (9.0 µL, 0.08 mmol) was added and the reaction mixture continued to stir at 40°C for 5 hr. LC-MS indicated 98% conversion to product. The crude reaction mixture was purified by column chromatography (12 g, 0-10% MeOH in DCM, gradient elution). Product fractions were concentrated in vacuo and the residue triturated with a mixture of diethyl ether, DCM and iso-hexane (2:1:2, 5.0 mL) to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(4-methylpiperazin-1-yl)acetamide (Example 9) as a light orange solid (26 mg, 47%): m/z 661 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.39 (3H, s), 2.69 - 2.60 (3H, bm), 2.88 - 2.73 (3H, bm), 3.17 - 2.95 (4H, bm), 5.39 (2H, s), 6.34 (1 H, s), 7.00 (1 H, d), 7.29 (1 H, d), 7.35 (2H, d), 7.45 (2H, d), 7.66 - 7.56 (3H, m), 7.98 (1 H, d), 8.37 - 8.28 (3H, m), 8.73 (1 H, s), 8.91 (1 H, s), 10.12 (1 H, s).

Example 10: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-(4-(2-methoxyethyl)piperazin-1-yl)acetamide

To a solution of N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (50 mg, 0.08 mmol) in DCM (1.0 mL), DMF (0.1 mL) and DIPEA (22 µl, 0.13 mmol) was added N-methoxyethyl piperazine (12.5 µl, 0.08 mmol). The reaction mixture was stirred at RT for 3 hr. LC-MS indicated 20% conversion to product. The reaction mixture was heated to 40°C and stirred for 12 hr. LC-MS indicated 78% conversion to product. A further portion of N-methoxyethyl piperazine (12.5 µL, 0.08 mmol) was added and the reaction mixture continued to stir at 40°C for 5 hr. LC-MS indicated 89% conversion to product. The crude reaction mixture was purified by column chromatography (12 g, 0-10% MeOH in DCM, gradient elution) to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(4-(2-methoxyethyl) piperazin-1-yl)acetamide (Example 10) as a light orange solid (45 mg, 73%): m/z 705 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO) δ: 1.27 (9H, s), 2.39 (3H, s), 2.46 - 2.48 (3H, m, obscured by DMSO), 2.57 - 2.50 (4H, m), 3.17 (2H, s), 3.23 (3H, s), 3.42 (2H, t), 5.39 (2H, s), 6.35 (1 H, s), 7.01 (1 H, d), 7.29 (1 H, d), 7.35 (2H, d), 7.43 (2H, d), 7.65 - 7.55 (3H, m), 7.93 (1 H, d), 8.36 - 8.30 (3H, m), 8.58 (1 H, s), 8.79 (1 H, s), 9.92 (1 H, s).

Example 11: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-(2-methoxyethylamino)acetamide

To a solution of N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (50 mg, 0.08 mmol) in DCM (1.0 mL), DMF (0.1 mL) and DIPEA (17 µL, 0.10 mmol) was added 2-methoxyethylamine (7.0 µL, 0.08 mmol). The reaction mixture was heated to 40°C and stirred for 12 hr. The crude reaction mixture was purified by column chromatography (12 g, 0-10% MeOH in DCM, gradient elution). Product fractions were concentrated in vacuo and the residue triturated with a mixture of diethyl ether, DCM and iso-hexane (2:1:2, 5.0 mL) to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(2-methoxyethylamino)acetamide (Example 11) as an off-white solid (6 mg, 11%): m/z 637 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.39 (3H, s), 2.71 (2H, t), 3.24 (3H, s), 3.33 (2H, m (obscured by DHO)), 3.40 (2H, t), 5.38 (2H, s), 6.35 (1 H, s), 7.01 (1 H, d), 7.27 (1 H, d), 7.36 (2H, d), 7.43 (2H, d), 7.64 - 7.57 (3H, m), 7.92 (1 H, m), 8.36 - 8.30 (3H, m), 8.59 (1 H, s), 8.79 (1 H, s).

Example 12: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-(dimethylamino)acetamide

To a solution of N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (50 mg, 0.08 mmol) in DCM (1.0 mL), DMF (0.1 mL) and DIPEA (17 µl, 0.1 mmol) was added dimethylamine (2.0M solution in THF) (41 µL, 0.08 mmol). The reaction mixture was heated to 40°C and stirred for 12 hr. The crude reaction mixture was purified by column chromatography (12 g silica, 0-10% MeOH in DCM, gradient elution). Product fractions were concentrated in vacuo and the residue triturated with a mixture of diethyl ether, DCM and iso-hexane (2:1:2, 5.0 mL) to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(dimethylamino) acetamide (Example 12) as an orange solid (18 mg, 35%): m/z 607 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.31 (6H, s), 2.39 (3H, s), 3.14 (2H, s), 5.39 (2H, s), 6.35 (1 H, s), 7.01 (1 H, d), 7.29 (1 H, d), 7.35 (2H, d), 7.44 (2H, d), 7.65 - 7.55 (3H, m), 7.94 (1 H, m), 8.38 - 8.28 (3H, m), 8.59 (1 H, s), 8.79 (1 H, s), 9.93 (1 H, s).

Example 13: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-(methylamino)acetamide

To a solution of N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (50 mg, 0.08 mmol) in DCM (1.0 mL), DMF (0.2 mL) and DIPEA (17 µL, 0.10 mmol) was added methylamine (2.0M solution in THF) (41 µl, 0.08 mmol). The reaction mixture was heated to 40°C and stirred for 12 hr. The crude reaction mixture was purified by column chromatography (12 g, 0-10% MeOH in DCM, gradient elution). Product fractions were contaminated with an impurity; the crude material was repurified by column chromatography (12 g, 0-10% MeOH in DCM, gradient elution) to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(methylamino)acetamide (Example 13) as a light brown solid (6 mg, 12%): m/z 593 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.32 (3H, s), 2.39 (3H, s), 3.28 (2H, s), 5.39 (2H, s), 6.35 (1 H, s), 7.01 (1 H, d), 7.27 (1 H, d), 7.35 (2H, d), 7.44 (2H, d), 7.63 - 7.55 (3H, m), 7.93 (1 H, m), 8.37 - 8.30 (3H, m), 8.59 (1 H, s), 8.80 (1 H, s).

Example 14: N-(4-((4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-((4-methoxybenzyl)(methyl)amino)acetamide

To a solution of N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) methyl)pyridin-2-yl)-2-chloroacetamide (Intermediate B) (50 mg, 0.08 mmol) in DCM (1.0 mL), DMF (0.2 mL) and DIPEA (17.5 µL, 0.10 mmol) was added N-(4-methoxybenzyl)-N-methylamine (15.5 µL, 0.09 mmol). The reaction mixture was stirred at 55°C for 12 hr. The crude reaction mixture was purified by column chromatography (12 g, 0-10% MeOH in DCM, gradient elution). Product fractions were concentrated in vacuo and the residue triturated with a mixture of diethyl ether, DCM and iso-hexane (2:1:2, 5.0 mL) to afford N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-((4-methoxybenzyl)(methyl)amino)acetamide (Example 14) as a white solid (7 mg, 11%): m/z 713 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.27 (9H, s), 2.25 (3H, s), 2.39 (3H, s), 3.22 (2H, s), 3.59 (2H, s), 3.72 (3H, s), 5.38 (2H, s), 6.35 (1H, s), 6.90 (2H, m), 7.01 (1H, m), 7.27 (3H, m), 7.35 (2H, m), 7.43 (2H, m), 7.64 - 7.55 (3H, m), 7.94 (1 H, m), 8.37 - 8.28 (3H, m), 8.58 (1 H, s), 8.79 (1 H, s), 9.97 (1 H, s).

Intermediate C: 1-(4-(2-(2-Aminopyridin-4-yl)ethoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1 H-pyrazol-5-yl)urea

tert-Butyl 4-(2-hydroxyethyl)pyridin-2-ylcarbamate (6)

To a solution of ethyl 2-(2-(tert-butoxycarbonylamino)pyridin-4-yl)acetate (5) ( WO 2007/089512 ) (10.0 g, 35.7 mmol) under nitrogen in THF (100 mL), at -78°C, was added DIBAL (1 M solution in THF, 71.3 mL, 71.3 mmol) over 1 hr. The reaction mixture was stirred at -78 to -60°C for 40 min and then warmed to -15°C over 1 hr. The solution was re-cooled to -78°C and treated with further DIBAL (1M solution in THF, 35 mL, 35.7 mmol). The mixture was allowed to warm to - 40°C and stirred for 1 hr. Water (10 mL) was added cautiously to quench the reaction followed by MgSO₄ (20 g) and the solids removed by filtration. The filtrate was concentrated to dryness under reduced pressure and the residue subjected to column chromatography (330 g), eluting with 65 % EtOAc in hexanes to give tert-butyl 4-(2-hydroxyethyl)pyridin-2-ylcarbamate (6) (6.00 g, 64%) as a yellow solid: m/z 239 (M+H)⁺ (ES⁺).

tert-Butyl 4-(2-(4-nitronaphthalen-1-yloxy)ethyl)pyridin-2-ylcarbamate (7)

To a solution of tert-butyl 4-(2-hydroxyethyl)pyridin-2-ylcarbamate (6) (6.00 g, 25.2 mmol) in THF (70 mL) was added sodium hydride (2.52 g, 63.0 mmol, 60 wt%) at 0°C. The bright yellow suspension was stirred for 20 min at 0°C before the addition of 1-fluoro-4-nitronaphthalene (4.81 g, 25.2 mmol) in a single portion. After stirring at RT for 2 hr, water (100 mL) was added followed by EtOAc (100 mL). The solid formed between the layers was collected by filtration and the organic phase was washed with saturated aq NaHCO₃ (100 mL), brine (100 mL) and dried. The volatiles were removed to give an orange solid. The solids were combined and triturated from MeOH (50 mL) to give tert-butyl 4-(2-(4-nitronaphthalen-1-yloxy)ethyl)pyridin-2-ylcarbamate (7) as a yellow solid (11.0 g, 98%): m/z 410 (M+H)⁺ (ES⁺).

tert-Butyl 4-(2-(4-aminonaphthalen-1-yloxy)ethyl)pyridin-2-ylcarbamate (8)

tert-Butyl 4-(2-(4-nitronaphthalen-1-yloxy)ethyl)pyridin-2-ylcarbamate (7) (5.20 g, 12.7 mmol) and iron mesh (4.30 g, 76 mmol) were suspended in a mixture of AcOH and EtOH (1:2, 120 mL). The suspension was placed in a pre-heated oil bath at 60°C and stirred rapidly until the reaction was judged to be complete by LC-MS. The mixture was cooled to RT, poured carefully onto saturated aq NaHCO₃ (1000 mL) and extracted with EtOAc (500 mL x 2). The combined organic layers were washed with further saturated aq NaHCO₃ (1000 mL), water (1000 mL), brine (1000 mL) and dried. This was filtered and evaporated to give tert-butyl 4-(2-(4-aminonaphthalen-1-yloxy)ethyl)pyridin-2-ylcarbamate (8) as a yellow oil (5.00 g, 95%): m/z 380 (M+H)⁺ (ES⁺).

tert-Butyl-4-(2-(4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)ethyl) pyridin-2-ylcarbamate (9)

To a suspension of CDI (3.00 g, 18.18 mmol) in DCM (15 mL) was added a solution of 3-tert-butyl-1-p-tolyl-1H-pyrazol-5-amine (4) ( WO 2000/043384 ) (4.17 g, 18.18 mmol) in DCM (40 mL) over 1.5 hr. After stirring at RT for 2 hr, a solution of tert-butyl 4-(2-(4-aminonaphthalen-1-yloxy)ethyl)pyridin-2-ylcarbamate (8) (3.00 g, 7.91 mmol) in DCM (15 mL) was added. After stirring overnight, the solution was diluted with MeOH (10 mL) and absorbed onto silica gel (30 g) and subjected to column chromatography (330 g) eluting with 30% to 100% EtOAc in iso-hexane and then 0% to 6% MeOH in EtOAc to give tert-butyl-4-(2-(4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)ethyl)pyridin-2-ylcarbamate (9) as a beige solid (4.20 g, 80%): m/z 635 (M+H)⁺ (ES⁺).

Intermediate C: 1-(4-(2-(2-Aminopyridin-4-yl)ethoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1 H-pyrazol-5-yl)urea

To a suspension of tert-butyl-4-(2-(4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido) naphthalen-1-yloxy)ethyl)pyridin-2-ylcarbamate (9) (1.35 g, 2.20 mmol) in DCM (10 mL) was added TFA (10 mL). After stirring at RT for 2 hr, the volatiles were evaporated and the residue was taken up in EtOAc (50 mL) and extracted with saturated aq NaHCO₃ (50 mL). The layers were separated; the organic was washed with brine (50 mL), dried and evaporated to give 1-(4-(2-(2-aminopyridin-4-yl)ethoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate C) as a pale pink solid (1.20 g, 100%): m/z 535 (M+H)⁺ (ES⁺).

Example 15: N-(4-(2-(4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) ethyl)pyridin-2-yl)-2-methoxyacetamide

To a suspension of 1-(4-(2-(2-aminopyridin-4-yl)ethoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1 H-pyrazol-5-yl)urea (Intermediate C) (35 mg, 0.065 mmol) in DCM (0.5 mL) was added DIPEA (23 µL, 0.131 mmol) and methoxyacetyl chloride (7 µL, 0.072 mmol). The mixture was stirred at RT, until judged to be complete by LC-MS; diluted with saturated aq NaHCO₃ (1.5 mL) and the layers were separated through a phase separator cartridge. The organics were collected, evaporated under reduced pressure and the residue subjected to SCX capture and release. The resulting residue was purified further by preparative RP HPLC to give N-(4-(2-(4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)ethyl)pyridin-2-yl)-2-methoxyacetamide (Example 15) as a white solid (5 mg, 13%): m/z 607 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.26 (9H, s), 2.37 (3H, s), 3.20 (2H, t), 3.37 (3H, s), 4.06 (2H, s), 4.38 (2H, t), 6.33 (1 H, s), 6.95 (1 H, d), 7.19 (1 H, dd), 7.33 (2H, m), 7.42-7.47 (3H, m), 7.54 (1 H, m), 7.59 (1 H, d), 7.87 (1 H, d), 8.12 (1 H, d), 8.18 (1 H, bs), 8.23 (1 H, d), 8.67 (1 H, s), 8.84 (1 H, s), 9.89 (1 H, s).

Example 16: N-(4-(2-(4-(3-(3-tert-Butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy) ethyl)pyridin-2-yl)-2-(2-methoxyethoxy)acetamide

To a suspension of 1-(4-(2-(2-aminopyridin-4-yl)ethoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate C) (35 mg, 0.065 mmol) in DCM (0.5 mL) was added DIPEA (23 µL, 0.131 mmol) and 2-(2-methoxy)ethoxyacetyl chloride (11 mg, 0.072 mmol). The mixture was stirred at RT, until judged to be complete by LC-MS; diluted with saturated aq NaHCO₃ (1.5 mL) and the layers were separated through a phase separator cartridge. The organics were collected, evaporated under reduced pressure and the residue subjected to SCX capture and release. The resulting residue was purified further by preparative RP HPLC to give N-(4-(2-(4-(3-(3-tert-butyl-1H-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)ethyl)pyrid in-2-yl)-2-(2-methoxyethoxy)acetamide (Example 16) as an off white solid (13 mg, 31%): m/z 651 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.26 (9H, s), 2.38 (3H, s), 3.21 (2H, t), 3.28 (3H, s), 3.49-3.51 (2H, m), 3.66-3.68 (2H, m), 4.13 (2H, s), 4.38 (2H, t), 6.34 (1 H, s), 6.95 (1 H, d), 7.19 (1 H, dd), 7.34 (2H, m), 7.41-7.48 (3H, m), 7.51-7.56 (1 H, m), 7.59 (1 H, d), 7.87 (1 H, d), 8.11-8.14 (1 H, dd), 8.20 (1 H, bs), 8.23-8.25 (1 H, dd), 8.55 (1 H, s), 8.75 (1 H, s), 9.83 (1 H, s).

Example 17: 1-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-(2-(2-(3-methylureido)pyridine-4-yl)ethoxy)naphthalen-1-ylurea

To a solution of 1-(4-(2-(2-aminopyridin-4-yl)ethoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate C) (50 mg, 0.094 mmol) in pyridine (1.0 mL) was added methyl isocyanate (5.34 mg, 0.094 mmol). The mixture was stirred at RT for 72 hr and the solvent was evaporated under reduced pressure. The resulting residue was triturated from MeOH (5.0 mL) to give 1-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-(2-(2-(3-methylureido)pyridine-4-yl)ethoxy)naphthalen-1-ylurea (Example 17) as an off white solid (7 mg, 13%): m/z 592 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.26 (9H, s), 2.38 (3H, s), 3.13 (2H, t), 3.31 (3H, s, obscured by H₂O),4.32 (2H, t), 6.34 (1 H, s), 6.93 (1 H, d), 7.34 (2H, m), 7.40-7.48 (6H, m), 7.53-7.57 (1 H, m), 7.61 (1 H, d), 7.81-7.83 (2H, d), 7.86-7.89 (1 H, d), 8.02-8.04 (1 H, dd), 8.55 (1 H, s), 8.75 (1 H, s).

Example 18: 1-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-(2-(2-ureidopyridine-4-yl)ethoxy)naphthalen-1-ylurea

To a solution of 1-(4-(2-(2-aminopyridin-4-yl)ethoxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea (Intermediate C) (50 mg, 0.094 mmol) in pyridine (1.0 mL) was added trichloroacetylisocyanate (12 µL, 0.103 mmol). The mixture was stirred at RT until judged to be complete by LC-MS and solvent was evaporated under reduced pressure. The resulting residue was subjected to SCX capture and release and triturated from DCM (10 mL) to give 1-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-(2-(2-ureidopyridine-4-yl)ethoxy)naphthalen-1-ylurea (Example 18) as an off white solid (25 mg, 44%): m/z 578 (M+H)⁺ (ES⁺). ¹H NMR (400MHz, DMSO-d₆) δ: 1.26 (9H, s), 2.38 (3H, s), 3.12 (2H, t), 4.35 (2H, t), 6.34 (1 H, s), 6.94-6.99 (2H, m), 7.19 (1 H, dd), 7.33-7.35 (2H, m), 7.41-7.50 (5H, m), 7.52-7.56 (1 H, m), 7.60 (1 H, d), 7.87 (1 H, d), 8.09-8.13 (2H, m), 8.54 (1 H, s), 8.75 (1 H, s), 9.08 (1 H, s).

Biological Testing

All compound examples demonstrated EC₅₀ values of less than 1 µM versus LPS-induced TNFα release in differentiated U937 cells (see below for assay details). Summaries of the properties of Example 1 established using both in vitro and in vivo assays are presented below.

In vitro testing for Example 1

Enzyme		Differentiated U937 cells		THP1 cells LPS-induced TNFα release
			MTT Assay
Alpha subtype	Gamma subtype		4, 24 h (10 µg/ml)
5.3	402	0.88		2.3

1.50% effective concentration relative to the effect of 10 µg/mL BIRB796 (as 100%). 2: no significant toxic effect observed in MTT assay

A description of these assays is as follows:

Enzyme inhibition assay

The enzyme inhibitory activity of compound was determined by fluorescence resonance energy transfer (FRET) using synthetic peptides labelled with both donor and acceptor fluorophores (Z-LYTE, Invitrogen). Briefly, recombinant, phosphorylated p38 MAPK gamma (MAPK12 :Millipore) was diluted in HEPES buffer, mixed with compound at desired final concentrations and incubated for two hr at RT. The FRET peptide (2 µM) and ATP (100 µM)were next added to the enzyme/compound mixture and incubated for one hr. Development reagent (protease) was added for one hr prior to detection in a fluorescence microplate reader. The site-specific protease only cleaves non-phosphorylated peptide and eliminates the FRET signal. Phosphorylation levels of each reaction were calculated using the ratio of coumarin emission (donor) over fluorescein emission (acceptor) with high ratios indicating high phosphorylation and low ratios, low phosphorylation levels. The percentage inhibition of each reaction was calculated relative to non-inhibited control, and the 50% inhibitory concentration (IC₅₀ value) then calculated from the concentration-response curve.

For p38 MAPK alpha (MAPK14: Invitrogen), enzyme activity was evaluated indirectly by determining activation/phosphorylation of the down-stream molecule, MAPKAP-K2. The p38 MAPK α protein was mixed with its inactive target MAPKAP-K2 (Invitrogen) and compound for two hr at RT. The FRET peptide (2 µM), which is a phosphorylation target for MAPKAP-K2, and ATP (10 µM) were then added to the enzymes/compound mixture and incubated for one hr. Development reagent was then added and the mixture incubated for one hr before detection by fluorescence completed the assay protocol.

LPS-induced TNF alpha release in U937 Cells: potency

U937 cells, human monocytic cell line, were differentiated to macrophage-type cells by incubation with phorbol myristate acetate (PMA; 100 ng/ml) for 48 to 72 hr. Where appropriate, cells were pre-incubated with final concentrations of compound for 2 hr. Cells were then stimulated with 0.1 µg/mL of LPS (from E. Coli: 0111 :B4, Sigma) for 4 hr, and the supernatant collected for determination of TNFα concentration by sandwich ELISA (Duo-set, R&D systems). The inhibition of TNFα production was calculated as a percentage of that achieved by 10 µg/mL of BIRB796 at each concentration of test compound by comparison with vehicle control. The 50% effective concentration (EC₅₀) was determined from the resultant concentration-response curve.

LPS-induced TNF alpha release in THP-1 Cells: potency

THP-1 cells, a human monocytic cell line, were stimulated with 1 µg/mL of LPS (from E. Coli; 0111:B4, Sigma) for 4 hr and the supernatant collected for determination of TNFα concentration by sandwich ELISA (Duo-set, R&D systems). The inhibition of TNFα production was calculated at each concentration by comparison with vehicle control. The 50% inhibitory concentration (IC₅₀) was determined from the resultant concentration-response curve.

MTT assay

Differentiated U937 cells were pre-incubated with compound for 4 hr in 5% FCS or 10% FCS for 24 hr and 72 hr. The supernatant was replaced with 200ul of new media and 10 µL of MTT stock solution (5 mg/ml) added to each well. After 1 hr incubation, the media were removed, 200 uL of DMSO added to each well and the plates were shaken lightly for 1 hr prior to reading the absorbance at 550nm.

The percentage loss of cell viability was calculated for each well relative to vehicle (0.5% DMSO)-treatment. Consequently an apparent increase in cell viability for drug treatment relative to vehicle is tabulated as a negative percentage.

In vivo testing for Example 1 LPS-induced neutrophilia in the mouse: duration of action

Non-fasted mice were dosed by the intra tracheal route with either vehicle, or the test substance at the time points ("pre-dose") indicated with respect to the start of LPS treatment. At T = 0, mice were placed into an exposure chamber and exposed to LPS. Eight hr after LPS challenge, animals were under anesthetized, the trachea cannulated and BALF extracted by infusing and withdrawing 1 mL of PBS into the lungs via a tracheal catheter. Total and differential white cell counts in the BALF samples were measured using a Neubaur haemocytometer. Cytospin smears of the BALF samples were prepared by centrifugation at 200 rpm for 5 min at RT and stained using a DiffQuik stain system (Dade Behring). Cells were counted using oil immersion microscopy.

The results are shown in Figures 1 and 2. Data for neutrophil numbers is reported as total and differential number (test substance relative to vehicle) of cells per mL of BALF, mean ± S.E.M. (n=8).

Summary

The biological studies in vitro show that the compound of Example 1 is a potent inhibitor of p38 MAP kinase subtypes alpha and gamma with good efficacy in an in vitro model of antiinflammatory activity (LPS-induced TNF alpha release from differentiated U937 cells and THP-1 cells). From the MTT results it may be concluded that the compound does not exhibit overt cellular toxicity at the concentrations used.

The biological studies in vivo show that the compound of Example 1 is effective in inhibiting LPS-induced neutrophil accumulation in an animal model, with a long duration of effect as shown by the significant inhibition even at 12 or more hr of pre-dosing.

Throughout the specification and the claims which follow, unless the context requires otherwise, the word 'comprise', and variations such as 'comprises' and 'comprising', will be understood to imply the inclusion of a stated integer, step, group of integers or group of steps but not to the exclusion of any other integer, step, group of integers or group of steps.

Claims (15)

1. A compound of formula (I)

   wherein R¹ is C_1-6 alkyl optionally substituted by a hydroxyl group;

   R² is H or C_1-6 alkyl optionally substituted by a hydroxyl group;

   R³ is H;

   Ar is a naphthyl or a phenyl ring either of which may be optionally substituted by one or more groups independently selected from C_1-6 alkyl, C_1-6 alkoxy, amino, and C_1-4 mono or di-alkyl amino;

   L represents -O(CH₂)_m- in which m is 0, 1 or 2;

   Q is selected from:

   a) a saturated or unsaturated, branched or unbranched C_1-10 alkyl chain, wherein at least one carbon (for example 1, 2 or 3 carbons) is replaced by a heteroatom selected from O, N, and S(O)_p, wherein said chain is optionally, substituted by one or more groups selected from oxo, halogen, an aryl group, a heteroaryl group and a heterocyclyl group, each aryl, heteroaryl or heterocyclyl group bearing 0 to 3 substituents selected from halogen, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, amino, and C_1-4 mono or di-alkyl amino, with the proviso that the atom linked directly to the carbonyl in -NR³C(O)- is not an oxygen or a sulfur atom; and

   b) a C_0-8 alkylC_5-6 heterocyclyl said heterocyclyl group comprising at least one heteroatom selected from O, N and S, and optionally substituted by one or two or three groups independently selected from halogen C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, amino, and C_1-4 mono or di-alkyl amino; and

   p is 0, 1 or 2;

   or a pharmaceutically acceptable salt or solvate thereof, including all stereoisomers and tautomers thereof.
2. A compound of formula (I) according to claim 1, wherein Ar is naphthyl.
3. A compound of formula (I) according to claim 1 or 2, wherein R¹ is tert-butyl.
4. A compound of formula (I) according to any one of claims 1 to 3, wherein R² is methyl.
5. A compound of formula (I) according to any one of claims 1 to 4, wherein R² is in the para position.
6. A compound of formula (I) according to any one of claims 1 to 5, wherein Q is selected from: -NR³C(O)CH₂OC_1-6 alkyl, -NR³C(O)CH₂O(CH₂)₂OCH₃, -NR³C(O)CH(CH³)OCH³, -NR³C(O)CH₂NHCH₃, -NR³C(O)CH₂NHCH₂CH₂OCH₃, -NR³C(O)CH₂SCH₃, - NR³C(O)NH₂, -NR³C(O)CH₂S(O)₂CH₃, -NR³C(O)NHC_1-7 alkyl, -NR³C(O)N(C_1-4 alkyl)C_1-5 alkyl, and -NR³C(O)CHN[(CH₂)₂OCH₃]_2.
7. A compound of formula (I) according to claim 6, wherein Q is selected from: -NHC(O)CH₂OCH₃; -NHC(O)CH₂O(CH₂)₂OCH₃; -NHC(O)CH(CH₃)OCH₃; -NHC(O)CH₂NHCH₃; -NHC(O)CH₂NH(CH₂)₂OCH₃, -NHC(O)CH₂SCH₃; -NHC(O)NH₂; -NHC(O)CH₂S(O)₂CH₃; -NHC(O)NHCH₃; -NHC(O)N(CH₃)₂; and -NHC(O)CHN[(CH₂)₂OCH₃]₂.
8. A compound of formula (I) according claim 1, wherein the compound is of formula (IA) wherein x is an integer from 1 to 6 and y is zero or an integer from 1 to 5 with the proviso that x + y is an integer from 1 to 6, for example x is 1 and y is 1; or a pharmaceutically acceptable salt or solvate thereof, including all stereoisomers and tautomers thereof; or wherein the compound is of formula (IB) wherein x is an integer from 1 to 6 and y is zero or an integer from 1 to 5, with the proviso that x + y is an integer from 1 to 6, for example x is 1 and y is 0; or a pharmaceutically acceptable salt or solvate thereof, including all stereoisomers and tautomers thereof.
9. A compound of formula (I) according to claim 1, wherein the compound is selected from:

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxyacetamide;

   1-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(3-methylureido)pyridin-4-yl)methoxy)naphthalen-1-yl)urea;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)tetrahydro-2H-pyran-4-carboxamide;

   (S)-N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxypropanamide;

   (R)-N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-methoxypropanamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(methylthio)acetamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-morpholinoacetamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(pyrrolidin-1-yl)acetamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(4-methylpiperazin-1-yl)acetamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthaten-1-yloxy)methyl)pyridin-2-yl)-2-(4-(2-methoxyethyl)piperazin-1-yl)acetamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(2-methoxyethylamino)acetamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(dimethylamino)acetamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-(methylamino)acetamide;

   N-(4-((4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)methyl)pyridin-2-yl)-2-((4-methoxybenzyl)(methyl)amino)acetamide;

   N-(4-(2-(4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)ethyl)pyridin-2-yl)-2-methoxyacetamide;

   N-(4-(2-(4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)ethyl)pyridin-2-yl)-2-(2-methoxyethoxy)acetamide;

   1-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-(2-(2-(3-methylureido)pyridin-4-yl)ethoxy)naphthalen-1-yl)urea; and

   1-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-(2-(2-ureidopyridin-4-yl)ethoxy) naphthalen-1-yl)urea;

   or a pharmaceutically acceptable salt or solvate thereof, including all stereoisomers and tautomers thereof.
10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 9, in combination with one or more pharmaceutically acceptable diluents or carriers.
11. A compound of formula (I) according to any one of claims 1 to 9 for use as a medicament.
12. A compound of formula (I) according to any one of claims 1 to 9 for use in the treatment or prevention of a condition selected from:

    a respiratory disorder such as COPD (including chronic bronchitis and emphysema), asthma, paediatric asthma, cystic fibrosis, sarcoidosis, idiopathic pulmonary fibrosis, allergic rhinitis, rhinitis or sinusitis,

    allergic conjunctivitis, conjunctivitis, allergic dermatitis, contact dermatitis, psoriasis, ulcerative colitis, inflamed joints secondary to rheumatoid arthritis or osteoarthritis,

    rheumatoid arthritis, pancreatitis, cachexia, inhibition of the growth and metastasis of tumours including non-small cell lung carcinoma, breast carcinoma, gastric carcinoma, colorectal carcinomas and malignant melanoma.
13. A compound of formula (I) for use according to claim 11 or claim 12 in combination with one or more other active ingredients.
14. Use of a compound of formula (I) according to any one of claims 1 to 9 for the manufacture of a medicament for the treatment or prevention of a condition selected from:

    a respiratory disorder such as COPD (including chronic bronchitis and emphysema), asthma, paediatric asthma, cystic fibrosis, sarcoidosis, idiopathic pulmonary fibrosis, allergic rhinitis, rhinitis or sinusitis,

    allergic conjunctivitis, conjunctivitis, allergic dermatitis, contact dermatitis, psoriasis, ulcerative colitis, inflamed joints secondary to rheumatoid arthritis or osteoarthritis, rheumatoid arthritis, pancreatitis, cachexia, inhibition of the growth and metastasis of tumours including non-small cell lung carcinoma, breast carcinoma, gastric carcinoma, colorectal carcinomas and malignant melanoma.
15. A compound of formula (I) for use according to claim 13 in the treatment or prevention of a respiratory disorder wherein the one or more other active ingredients are selected from steroids, beta agonists and xanthines.